A vesicular, immunogenic, non-pyrogenic complex is obtained from isolates of N. gonorrhoeae by cultivating N. gonorrhoeae on a suitable nutrient medium; harvesting and centrifuging the cells; extracting the cell pellet with TRITON X-100 at alkaline pH; suspending the solids remaining after extraction in an aqueous medium and precipitating in the cold with ethanol.
The instant invention relates to a novel vesicular, immunogenic, non-pyrogenic complex obtained from isolates of N. gonorrhoeae. More particularly, the instant invention relates to said immunogenic complex; to the process for obtaining said immunogenic complex from isolates of N. gonorrhoeae; and to compositions for the safe and effective administration of said immunogenic complex to mammalian species in order to provide protection against N. gonorrhoeae.
The instant invention is based upon applicant's discovery that a vesicular, immunogenic, non-pyrogenic complex may be obtained from isolates of N. gonorrhoeae by cultivating an isolate of N. gonorrhoeae on a suitable nutrient medium; harvesting and centrifuging the cells; extracting the cells with TRITON X-100 (polyoxyethylene octyl phenol, a non-ionic detergent) at alkaline pH; suspending the solids remaining after extraction in an aqueous medium and precipitating in the cold with ethanol; and releasing and removing RNA and DNA by nuclease digestion followed by diafiltration to remove the enzymes and degradation products. The material is then dispersed and solubilized by sonication and filtered through a 0.45 .mu.m polysulphone membrane. It is contemplated that immunogenically effective amounts of the complex of this invention in suitable formulations will be administered as a vaccine against N. gonorrhoeae infection.
Procedures for cultivating isolates of N. gonorrhoeae and for producing the immunogenic complex of this invention therefrom are described in detail below.